117-1Immunological testing• Monoclonal antibodies• Serology• Quantifying antigen – antibody reactions17-2Perspective 17.1Monoclonal Antibodies217-3Serology• Antibodies• Antibodies detect and identify antigens17-4Quantifying antigen – antibody reactions• Seroconversion or rise in titer• Serial dilutions317-5Figure 17.2 - Quantitation of immunologic tests17-6Precipitation reactions• Immunodiffusion• Immunoelectrophoresis417-7Figure 17.3Precipitation reaction17-8Figure 17.4Immunodiffusion517-9Figure 17.6Immunoelectrophoresis17-10Agglutination reactions• Direct agglutination• Indirect agglutination• Hemagglutination617-11Direct agglutination• Cross – linking and lattice formation• Antibodies react with particulate antigens (red blood cells, bacteria, fungi)• Visible clumps • Estimate amount of antibody17-12Indirect agglutination• Soluble antigen is coated onto particles (red bood cells, latex beads)• Allow for visible clumps (agglutination)717-13Figure 17.7 - Agglutination reaction17-14Immunofluorescence tests• Direct fluorescent antibody test• Indirect fluorescent antibody test817-15Figure 17.8 - Direct andindirect fluorescent antibody test17-16Antigen – antibody assays• Radioimmunoassay (RIA)• Enzyme – linked immunosorbant assay (ELISA)• Western blot917-17Radioimmunoassay (RIA)• Competitive inhibition assay• Measure antigen or antibody• Ex. Measure small amounts of hormones or drugs in a clinical sample• Ex. Measure small amounts of IgE antibody (radioallergosorbent test)17-18Enzyme – linked immunosorbant assay (ELISA)• Color reaction assay• Indirect ELISA• Direct ELISA1017-19Figure 17.9 - Indirect ELISA17-20Figure 17.1 - Direct ELISA1117-21Figure 17.1Western blot17-22Complement fixation test• Measures the binding of complement by an antigen –antibody interaction• Indicator system determine positive or negative reactions1217-23Figure 17.1Complementfixation test17-24Neutralization test• Antibody bind to specific antigen (virus, toxin)• Antibody – antigen complex prevents antigen from binding (neutralization)1317-25Cellular immunology test• Identification of subsets of lymphocytes• Lymphocyte response to mitogens• Cytoxic T – cell function• Cell – mediated immunity to infectious